Difference between revisions of "Part:BBa K3282001"

Revision as of 10:46, 16 October 2019

C. metallidurans lead transport protein - pbrT

This part contains coding sequence of Cupriavidus metallidurans CH34 lead transport protein (pbrT, Gene ID: 3170370, Uniprot: Q58AJ4) codon-optimized for Escherichia coli Nissle 1917. pbrT is a lead(II) uptake protein containing a cytochrome domain and a transmembrane metal transport domain [1]. It belongs to the ILT (Iron Lead Transporter) family. This protein is a part of the lead (Pb) resistance operon in plasmid pMOL30 [2]. Expression of pbrT leads to uptake of lead into the cytoplasm to reduce interaction of free Pb(II) with side chains of membrane and periplasmic proteins, which would cause extensive cellular damage [3]. The part was evaluated in the composite parts BBa_K3282005 and BBa_K3282007 expressed in pUC19 backbone (part---). The composites have the same RBS (BBa_J61109), terminator (BBa_B0012) with two different promoters T7 (BBa_I719005) and Tac (BBa_K3282006). The four composite parts were tested to determine their Lead uptake capability.

References

1. Taghavi, S., Lesaulnier, C., Monchy, S., Wattiez, R., Mergeay, M., & Lelie, D. V. D. (2008). Lead(II) resistance in Cupriavidus metallidurans CH34: interplay between plasmid and chromosomally-located functions. Antonie Van Leeuwenhoek, 96(2), 171–182. doi: 10.1007/s10482-008-9289-0.

2. Hobman, J. L., Julian, D. J., & Brown, N. L. (2012). Cysteine coordination of Pb(II) is involved in the PbrR-dependent activation of the lead-resistance promoter, PbrA, from Cupriavidus metallidurans CH34. BMC Microbiology, 12(1), 109. doi: 10.1186/1471-2180-12-109

3. Borremans, B., Hobman, J. L., Provoost, A., Brown, N. L., & van Der Lelie, D. (2001). Cloning and functional analysis of the pbr lead resistance determinant of Ralstonia metallidurans CH34. Journal of bacteriology, 183(19), 5651–5658. doi:10.1128/JB.183.19.5651-5658.2001

Sequence and Features